Our overall objective is to understand mechanisms of active transport and energy transduction in molecular terms. We plan to test pure isolated subunits of the energy transducing F1 -F ATPase complex of Escherichia coli by the "mixed reconstitution technique." A mixture of wild type and mutant subunits will be reconstituted in order to localize the cause of various disorders of oxidative phosphorylation to a particular subunit of the complex. We also plan to isolate transport carrier proteins from Escherichia coli and to reconstitute them in phospholipid vesicles. We shall continue our investigation of how ATP energy is used to drive active transport for shock-sensitive, binding protein systems. We shall explore the molecular mechanism whereby ATP increases the permeability of transformed but not untransformed mouse fibroblasts in culture. A particular protein kinase and phosphoprotein phosphatase will be continued, since it shows interesting differences between transformed and untransformed cells. The ATP permeabilization technique will be used to study control of glycolysis, the hexose monophosphate shunt and macromolecular synthesis in intact tumor cells, and the use of ATP to introduce normally impermeant drugs will be explored.